BACKGROUND:Diabetic nephropathy is an important cause of end-stage renal disease,and intestinal barrier damage plays an important role in the occurrence and development of diabetic nephropathy. OBJECTIVE:To observe the protective effect of human umbilical cord mesenchymal stem cells on the intestinal barrier in rats with diabetic nephropathy. METHODS:Thirty 8-week-old male SD rats were randomly assigned to healthy control group,model group and human umbilical cord mesenchymal stem cell group,with 10 rats in each group.Rats in the human umbilical cord mesenchymal stem cell group were injected with 1×106 human umbilical cord mesenchymal stem cells through the tail vein once a week for 4 weeks after the model establishment of diabetic nephropathy.Rats in the healthy control group and the model group were injected with an equal volume of PBS at the same time.1 week after the last injection,the histomorphological changes in the kidney and colon were observed under a light microscope.The expressions of ZO-1 and Occludin in the colon tissue of rats were detected by immunohistochemistry.Serum D-lactic acid and lipopolysaccharide levels were detected by ELISA.In addition,the distribution of human umbilical cord mesenchymal stem cells labeled with DiR dye in rats was observed by in vivo imaging system.The expression of human mesenchymal stem cell surface marker antigens CD44 and CD90 in colon tissue was detected by immunohistochemistry. RESULTS AND CONCLUSION:(1)Compared with the model group,human umbilical cord mesenchymal stem cell transplantation significantly inhibited the increase of urea nitrogen,serum creatinine,24-hour urine protein level and urinary albumin/creatinine ratio in diabetic nephropathy rats(all P<0.05).(2)The expression of human mesenchymal stem cell surface markers CD44 and CD90 was found in the colon of diabetic nephropathy rats.(3)Compared with the healthy control group,the expression levels of tight junction proteins Occludin and ZO-1 in the colon tissue of the model group were significantly reduced,while the expressions of Occludin and ZO-1 were significantly increased after treatment with human umbilical cord mesenchymal stem cells.(4)Compared with the model group,human umbilical cord mesenchymal stem cell transplantation significantly reduced serum D-lactic acid and lipopolysaccharide levels in diabetic nephropathy rats.(5)The results suggest that human umbilical cord mesenchymal stem cells may protect the intestinal barrier function by enhancing the expression of intestinal tight junction proteins in diabetic nephropathy rats.

OBJECTIVE: To investigate the clinical and genetic characteristics of a patient with STISS syndrome due to variant of PSMD12 gene. METHODS: Clinical data and result of genetic testing of a patient who was admitted to Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine on October 4, 2020 were analyzed, together with a review of relevant literature. RESULTS: The patient was found to harbor a heterozygous c.601C>T (p.Arg201*) nonsense variant of the PSMD12 gene, which was unreported previously. Clinically, the height of the patient has differed significantly from reported in the literature. An extremely rare case of STISS syndrome due to variant of the PSMD12 gene has been diagnosed. CONCLUSION Whether the severely short stature is part of the clinical spectrum for PSMD12 gene variants needs to be further explored, and the efficacy and safety of growth hormone therapy has yet to be determined.
Child ; Humans ; China ; Dwarfism ; Genetic Testing ; Heterozygote ; Syndrome

OBJECTIVE: To investigate the effect of Porphyromonas gingivalis (Pg) infection on IFNGR1 palmitoylation and biological behaviors of esophageal squamous cell carcinoma (ESCC) cells and the clinical implications. METHODS: The expression levels of IFNGR1 protein in ESCC cell lines KYSE30 and KYSE70 were detected using Western blotting at 24 and 48 h after Pg infection, and 2-BP was used to detect IFNGR1 palmitoylation in the cells. KYSE70 cells with wild-type IFNGR1 (IFNGR1-WT cells) and with IFNGR1-C122A palmitoylation site mutation induced by site-specific mutagenesis (IFNGR1-C122A cells) were both infected with Pg, and the changes in palmitoylation of IFNGR1-C122A were analyzed using immunofluorescence and Click-iT assays. The changes in proliferation, migration and invasion ability of the infected cells were evaluated using plate cloning assay, scratch assay and Transwell assay, and IFNGR1 co-localization with lysosomal marker LAMP2 was dected using immunofluorescence assay. Immunohistochemistry was used to detect Pg infection and IFNGR1 protein expression in 50 ESCC tissues, and their correlation with the clinicopathological characteristics and survival outcomes of the patients was analyzed. RESULTS: Pg infection down-regulated the protein expression of IFNGR1 in ESCC and promoted IFNGR1 palmitoylation at site 122. In IFNGR1-WT cells, Pg infection significantly enhanced cell proliferation, migration and invasion (P < 0.05). Similarly, Pg also significantly promoted proliferation, migration and invasion of IFNGR1-C122A cells, but to a lesser extent as compared with the wild-type cells (P < 0.05). Immunofluorescence assay showed that Pg and ZDHHC3 promoted IFNGR1 degradation within the lysosome. Immunohistochemical studies of the ESCC tissue samples showed a negative correlation between IFNGR1 and Pg expression, and a reduced IFNGR1 expression was correlated with a poorer survival outcome of the patient. CONCLUSION Pg infection enhances IFNGR1 palmitoylation to promote progression of ESCC, and elimination of Pg and inhibiting IFNGR1 palmitoylation may effectively control ESCC progression.
Humans ; Esophageal Neoplasms ; Porphyromonas gingivalis ; Lipoylation ; Esophageal Squamous Cell Carcinoma ; Lysosomes

Objective: To investigate the effect of Porphyromonas gingivallis ( Pg) infection of esophageal cancer cells on the polarization of tumor associated macrophages (TAMs) and functional changes. Methods: The secretion of tumor⁃related cytokines in the supernatant of Pg infected and uninfected esophageal squamous cell carcinoma (ESCC) cells was detected by ELISA. A co⁃culture model of ESCC cells and macrophages in vitro was established ,and the changes of TAMs surface markers were detected by qPCR , cellular immunofluorescence and flow cytometry. Cytokines secreted by TAMs after co⁃culture were detected by ELISA. ESCC cells were cultured using conditioned medium of co⁃cultured TAMs , and the effects of TAMs on the proliferation , migration and invasion of esophageal cancer cells were evaluated by CCK⁃8 , Wound⁃healing assay and Transwell assay. Results: The expression quantity of IL⁃6 and IL⁃10 of Pg⁃infected ESCC cells increased (P < 0. 01) . The contents of CD163 and CD206 on the surface of TAMs co⁃cultured with Pg⁃infected ESCC cells increased (P < 0. 001) . The cytokines IL⁃6 and IL⁃10 secreted by TAMs co⁃cultured with Pg⁃infected ESCC cells relatively increased (P < 0. 01) . TAMs co⁃cultured with Pginfected ESCC cells were able to enhance ESCC cells proliferation , migration and invasion (all P < 0. 05) . Conclusion Pg infection of ESCC cells can induce the secretion of cytokines , remodel TAMs to polarize toward the M2type immunosuppressive phenotype , thereby promoting the malignant biological behavior of ESCC cells. This study provides data support for the etiology of esophageal cancer and potential target molecules for clinical immunotherapy targeting TAMs.

Objective : To explore the relationship between Porphyromonas gingivalis(Pg) and mitophagy in esopha- geal cancer cells，and to explore new therapeutic targets for esophageal cancer． Methods : ① Western blot was used to detect the phosphorylation of unc-51-like authophagy activating kinase1 (ULK1) in mitochondria of the Pg infected cells and immunohistochemical method was used to detect the correlation between the expression of Pg and the phosphorylation status of ULK1 in esophageal cancer tissues. ② Western blot，ICC and ELISA were used to de- tect the transfer of nucleotide blinding domain and leucine rich repeat containing family member X1 (NLRX1) from cytoplasm to mitochondria，mitophagy，and the secretion levels of interleukin ( IL) -6 and reactive oxygen species (ROS) under Pg infection. ③ Pg colonization in esophageal tissues of mice in each group was detected by qPCR and Pg colonization in esophageal squamous epithelial cells of mice by RNAscope. Results : Compared with the un- treated group，the phosphorylation level of mitochondrial ULK1 (P＜0．01) ，NLRX1 expression (P＜0. 001) and mitophagy (P＜0. 001) of esophageal cancer cells increased after Pg infection．Compared with the control group， the combined intervention group could inhibit Pg colonization in esophageal tissue and esophageal squamous epithe- lial cells of mice (P＜0. 001) ． Conclusion Pg promotes the translocation of NLRX1 from cytoplasm to mitochon- dria by up-regulating the phosphorylation level of ULK1 in the mitochondria of esophageal cancer cells，and then induces mitophagy，leading to the reduction secretion of IL-6 and ROS，and ultimately maintaining Pg colonization.

Objectives To study the effect of cadmium (Cd) on the proliferation and apoptosis of mouse spermatocyte (GC-2 spd) cells and explore the underlying molecular mechanism. Methods GC-2 spd cells were cultured with 0, 5, 10, 15, 20 and 30 μM CdCl2, respectively, for 24 hours. The cell viability and IC50 of Cd were estimated based on CCK-8 data. The apoptosis of GC-2 spd cells and cellular concentration of ROS were analyzed by flow cytometry after treatment of the cells with different concentrations of CdCl2 (0, 5, 10 μM) for 24 hours. The expression levels of JNK/c-Jun signaling pathway regulatory proteins, pro-apoptotic factor Bax and anti-apoptotic factor Bcl-2, were determined by Western blot. Results Cd inhibited the proliferation of GC-2 spd cells with IC50 value of 12.99 μM, 95% CI [11.95, 14.00]. Exposure to 5 and 10 μM CaCl2 resulted in increases in apoptosis and cellular ROS generation in a dose-dependent manner, which was statistically significant compared with the control (P < 0.05). Although there was no difference in the expression level of c-Jun (P > 0.05), the phosphorylation level of JNK and c-Jun in Cd group was highly increased as compared to the control (P < 0.05). In addition, Cd exposure significantly increased the expression of Bax protein but decreased the expression Bcl-2 protein (P < 0.05). Conclusions Cadmium induces GC-2 spd cell apoptosis by increasing concentration of ROS and regulating the JNK/c-Jun signaling pathway.

OBJECTIVE：To shar e the experienc e of pharmaceutical care in Wuhan Jinyintan Hospital （herein after refers to “our hospital ”）under the condition of novel coronavirus pneumonia （COVID-19）epidemic，and to provide reference for other hospitals to deal with public health emergencies. METHODS ：The situation of pharmaceutical care in our hospital under the condition of COVID- 19 epidemic was summarized and shared ，including the epidemic prevention and control management （regional division ，disinfection management ，pharmacy personnel training ），supply of drugs and disinfection products ，the monitoring and education of rational drug use by information technology. RESULTS ：The pharmacy department of our hospital divided the activity scope into clean area ，potential pollution risk area ，semi pollution area ，and implement different disinfection management. All pharmacists received training ，involving personal health protection ，prevention and control knowledge of COVID-19，health status monitoring ，etc. For supply and guarantee of drugs and disinfectants ，the epidemic drug list of our hospital was formulated ，drugs and disinfectants were purchased accurately and stored in a standardized way. 24 h response telephone was set up in the clinical pharmacy room to receive consultation from clinicians on drug use at any time. The drugs mentioned in the COVID- 19 diagnosis scheme were compared in terms of the mechanism of action and the medication of special populations to form a tablet ，so as to help clinical rational choice treatment drug. CONCLUSIONS ：The pharmaceutical care in the designated hospital of COVID- 19 is a professional and complicated work ，involving a wide range of aspects. Pharmacy department must respond actively and adjust the strategy in time so as to play an important role in improving the ability of medical treatment.

Objective:To study the new mechanism of Xuebijing injection improving the function of pulmonary vascular barrier from the perspective of claudin-5 protein.Methods:Acute lung injury (ALI) model was induced by hydrogen sulfide (H 2S) exposure. ① In vivo study: Sprague-Dawley (SD) rats were divided into control group, H 2S exposure group (exposure to 300×10 -6 H 2S for 3 hours), Xuebijing control group (Xuebijing injection 4 mL/kg, twice a day, for 3 days), and Xuebijing intervention group (H 2S exposure after pretreatment of Xuebijing injection) according to random number method, with 6 rats in each group. At different time points (0, 6, 12 and 24 hours) after the model was made successfully, the total protein content in plasma and bronchoalveolar lavage fluid (BALF) of rats were detected respectively, and the pulmonary permeability index (PPI) was calculated (PPI = protein content in BALF/protein content in plasma), lung dry/wet weight ratio (W/D) was detected, and claudin-5 mRNA expression in lung tissue was measured by real time-polymerase chain reaction. ② In vitro test: human pulmonary microvascular endothelial cells (HPMECs) were divided into blank control group, NaHS treatment group (co-incubated with 500 μmol/L NaHS for 12 hours), Xuebijing control group (2 g/L Xuebijing injection for 24 hours), and Xuebijing intervention group (2 g/L Xuebijing injection pre-treated for 24 hours, then co-incubated with 500 μmol/L NaHS for 12 hours). The HPMECs claudin-5 protein expression and monolayer permeability changes were measured at different co-incubation time (1, 3, 6, 12 and 24 hours) by Western Blot and fluoresceinsodium. Results:① In vivo study: compared with the control group, the lung W/D ratio increased significantly at 6 hours and peaked at 12 hours after H 2S exposure in rats (4.67±0.11 vs. 4.26±0.06, P < 0.01). The expression of claudin-5 mRNA in lung tissue was significantly decreased, which was 89% of control group 6 hours after exposure ( P < 0.01). The total protein content in BALF and PPI at 12 hours after exposure were significantly higher than those in the control group [total protein content (mg/L): 262.31±14.24 vs. 33.30±3.09, PPI: (11.72±0.57)×10 -3 vs. (1.21±0.08)×10 -3, both P < 0.01], while the results in Xuebijing intervention group were significantly decreased [total protein content (mg/L): 153.25±7.32 vs. 262.31±14.24, PPI: (5.79±0.23)×10 -3 vs. (11.72±0.57)×10 -3, both P < 0.01]. ② In vitro test: compared with the blank control group, after incubating HPMECs with NaHS, the permeability of monolayer endothelial cells gradually increased, reaching the highest level in 12 hours, about twice of that in the blank control group, while claudin-5 protein expression decreased to the lowest level at 12 hours (claudin-5/β-actin: 0.42±0.03 vs. 1.03±0.05, P < 0.01). After intervention with Xuebijing, the permeability of endothelial cells was significantly improved (fluorescence intensity of fluorescein sodium: 1.46±0.10 vs. 1.89±0.11, P < 0.01), and the decrease of claudin-5 protein was reduced (claudin-5/β-actin: 0.68±0.04 vs. 0.38±0.03, P < 0.01). Conclusion:Xuebijing injection may improve pulmonary vascular barrier function in ALI by upregulating claudin-5 expression.

OBJECTIVE：To study the mechanism of Periplaneta americana extract degreasing cream and CⅡ-3（shorted for “degreasing cream ”and“CⅡ-3”）reversing the multi-drug resistance of human HepG 2/ADM cells. METHODS ：MTT assay was used to investigate the toxicity effects of different concentrations of sorafenib （positive control ），degreasing cream and C Ⅱ-3 on HepG2/ADM cells ，then IC 20 was calculated. The experiment was divided into sensitivity drug ，drug-resistance group ，sorafenib group，degreasing cream group and C Ⅱ-3 group. HepG 2 cells were included in sensitivity group ，and HepG 2/ADM cells were included in the latter 4 groups. Sensitivity group and drug-resistance group were treated with routine medium ，and other 3 groups were treated with relevant medicine （IC20 as drug concentration ）. The content of ADM in HepG 2/ADM cells was determined by Laser scanning confocal microscopy. The expression of apoptosis-related protein as Bcl- 2 and Cleaved-Caspase- 9 p37 were detected by Western blotting assay. RT-qPCR and immunocytochemistry were adopted to detect mRNA and protein expressions that related to multidrug resistance [P-gp （expression produce of MDR1 gene），LRP，BCRP] and that related to enzyme-mediated multidrug resistance pathway （GST-π and Topo Ⅱ）. RESULTS ：The IC 20 of degreasing cream ，CⅡ-3 and sorafenib were （2.40±0.16）， （200.44±27.52），（18.00±1.82）μg/mL，respectively. Compared with sensitivity group ，the protein expressions of Bcl- 2，P-gp， LRP，BCRP and Topo Ⅱ，the mRNA expressions of MDR 1， LRP，BCRP and GST-π were increased significantly in drug resistance group （P＜0.05 or P＜0.01）. Compared with @qq.com drug-resistance group ，the mRNA and protein expression of MDR1 mRNA and LRP ，BCRP，GST-π were significantly decreased in degreasing cream group and C Ⅱ-3 group（P＜ 0.05 or P＜0.01）；the protein expression of Bcl- 2 and the mRNA expression of Topo Ⅱ were significantly decreased （P＜0.01）， while the protein expression level of Cleaved-Caspase- 9 p37 was significantly increased in C Ⅱ -3 group （P＜0.05）. CONCLUSIONS：Degreasing cream and C Ⅱ-3 can reverse multidrug resistance of HepG 2/ADM cells by reducing drug efflux ， promoting cell apoptosis ，reducing the mRNA and protein expression of multi-drug resistance gene as well as gene in enzyme-mediated multi-drug resistance pathway. The effect of C Ⅱ-3 is better than that of degreasing cream.

Objective To explore the effect of fish oil fat emulsion as perioperative nutritional support on patients with liver cirrhosis and portal hypertension.Methods Randomized controlled clinical trial was performed between September 2011 and September 2017 in patients with liver cirrhosis and portal hypertension who underwent pericardial devascularization and splenectomy.Hypocaloric total parenteral nutritional support (TPN) started from the first day after the operation for 5 consecutive days.Patients were divided into experimental group and control group according to thetype of fat emulsion used.43 patients in experimental group were applied for fish oil fat emulsion injection (10％ Omegaven) + medium long chain structure fat emulsion (20％STG) and 42 patients in control group were applied for medium long chain structure fat emulsion (20％STG).Liver function (total bilirubin and alanine aminotransferase),nutrition index (serum albumin and prealbumin),inflammatory mediators (TNF-α,IL-6 and IL-10) were measured before and after the operation,and the clinical outcomes were observed.Results There was no statistically significant difference in liver function and nutritional indices between the experimental group and the control group (P＞0.05).The inflammatory mediators like TNF-u,IL-6 and IL-10 on the first day after surgery were significantly higher than those before surgery in both groups [experiment group:(225.54±54.78) vs.(61.49±16.47),(74.94±6.36) vs.(39.84±2.77),(77.53±11.4) vs.(46.05±6.13) ng/L;control group:(229.26±62.15) vs.(63.48±13.76),(77.23±7.83)vs.(40.64±3.34),(73.89±7.97)vs.(44.88±5.72) ng/L;P＜ 0.01].With the progress of time,the proinflammatory factors like TNF-α and IL-6 decreased after the operation and the range of decrease was higher in experiment group than in control group [d4-d 1:(-56.88± 31.63) vs.(-35.96±20.02),(-13.52±5.20) vs.(-6.38±2.84) ng/L;d7-d1:(-150.67±42.58) vs.(-132.79±53.35),(-27.04±8.97) vs.(-20.85±6.38) ng/L;P＜ 0.05].The range of increase in anti-inflammatory media IL-10 was higher in experiment group than in the control group (d4-d1:(14.22±13.08) vs.(5.64±3.58) ng/L;d7-d1:(17.78±5.58) vs.(-37.96±11.43) ng/L;P＜0.05).The incidence of grade Ⅲ complications and total complications (4.7％ vs.21.4％,23.3％ vs.45.2％) and hospitalization time [(10.12 ±1.48) vs.(12.33±2.04) d] in the experimental group were significantly lower than those in the control group (P＜0.05).Conclusions In patients with liver cirrhosis and portal hypertension,perioperative nutritional support of fish oil fat emulsion can reduce systemic inflammatory response and operative complications and promote rapid recovery through its two-way regulation of inflammatory mediators.